LOCUS       sequence-113840-         569 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses FokI-dCas9 and gRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 569)
  AUTHORS   Nakagawa Y, Sakuma T, Sakamoto T, Ohmuraya M, Nakagata N, Yamamoto T
  TITLE     Production of knockout mice by DNA microinjection of various 
            CRISPR/Cas9 vectors into freeze-thawed fertilized oocytes.
  JOURNAL   BMC Biotechnol. 2015 May 22;15(1):33.
  PUBMED    25997509
REFERENCE   2  (bases 1 to 569)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "BMC 
            Biotechnol."; date: "2015-05-22"; volume: "15(1)"; pages: "33"
COMMENT     Sequence Label: pX330A_FokI-1x4
FEATURES             Location/Qualifiers
     source          1..569
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     complement(39..59)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(39..57)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(91..113)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    102..118
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(126..132,133..150,151..156))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    171..192
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     enhancer        244..529
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 ttaccgggcc ccccctcgag gtcctccagc ttttgttccc tttagtgagg gttaattgcg
       61 cgcttggcgt aatcntggnc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
      121 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
      181 taactcacat taattgcgtt gcgctcactg cccgctttcc accggtggtc tcttagaggt
      241 acccgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
      301 attgacgtca atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
      361 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
      421 tgacgtcaat gacggnaaat ggcccgcctg gcattgtgcc cagtacatga ccttatggga
      481 ctttcctact tggnagtaca tctacgtatt agtcatcgct attaccatgg tcgagganag
      541 ccccacgttc tgcttcactc tccccatct
//
